Bacteriophages as recognition and identification agents
AssigneeThe Board of Trustees of the University of Illinois
InventorsMarius C. Teodorescu, Alexandre M. Gaspar
Life of this patent
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Abstract
Bacteriophages are employed as agents for recognition and identification of molecules and cellular materials, using their ability to recognize their bacterial host, by coating them with antibodies or by selecting them to perform in a manner analogous to antibodies. Visibility for identification is effected by incorporating a fluorescent agent, a radioisotope, a metal, an enzyme, or other staining material. The bacteriophage are prepared so as to bind to the molecular or cellular material through either the tail or head segment of the bacteriophage.
Claims6
A method for preparing a bacteriophage binding through a head segment to molecular or cellular materials, comprising the steps of:
- (a) mixing the bacteriophage, having both head and tail segments, with bacteria;
- (b) irrading the mixture with ultraviolet light to effect mutation of the bacteriophage;
- (c) nultiplying the mutant bacteriophage in the presence of bacteria binding through a tail segment to molecular of cellular material;
- (d) purifying the population of nutant bacteriophage;
- (e) binding the pruified mutant bacteriophage to the cellular material or to the molecular material;
- and
- (f) multiplying the bound mutant bacteriophage in bacteria.
A method for preparing a selected bacteriophage binding through a tail segment to molecular or cellular materials, comprising the steps of:
- (a) injecting molecular or cellular material into an animal host for the preparation of antibodies thereto;
- (b) recovering and purifying said antibodies;
- (c) coupling the purified antibodies to a solid phase;
- (d) binding the coupled antibodies with bacteria characterized as being resistant to a selected bacteriophage;
- (e) growing and harvesting the bacteria bound to said antibodies;
- (f) irradiating selected bacteriophage binding through a tail sesgment to molecular of cellular material with ultraviolet light;
- (g) separately irradiating a strain of bacteria, sensitive to the selected bacteriophage, with ultraviolet light;
- (h) mixing the irradiated bacteriophage of step (f) with the irradiated bacteria of step (g);
- (i) mixing approximately equal portions of the products of steps (e) and (h) under conditions for the development of plaques, whereby mutant bacteriophages are generated and yield clear plaques;
- and
- (j) recovering the mutant bacteriophage from the clear plaques and multiplying it in the bacteria of step (e), whereby there is obtained a mutant bacteriophage capable of binding to said molecular or cellular materials.
Dependencies are inferred from the claim text.